Preparation and identification of pseudovirus control samples for Nipah virus N gene nucleic acid detection

  Objective   To prepare safe and stable control samples for Nipah virus （NiV） molecular detection by recombinant pseudovirus containing N gene fragment of NiV.

  Methods   The nucleotide sequence containing NiV N gene fragment was synthesized and inserted into CD513b lentiviral vector, and the recombinant lentiviral vector and the lentivirus-packaging vector were co-transfected into HEK293T cells. The pseudoviral particles were concentrated and purified by ultracentrifugation, and the particle morphology was observed by transmission electron microscopy. The function of the pseudovirus was verified by infection experiment, and the linearity, stability and homogeneity were conducted by qRT-PCR.

  Results  The packaged and purified pseudoviral particles had a high concentration, with a diameter of 100−200 nm, and obvious G protein spikes were visible. The GFP signal of infection experiment was strong and the storage stability was good at different temperatures, and the coefficient of variation of homogeneity detection was less than 1%.

  Conclusion  The control samples of pseudovirus containing NiV N gene fragment were successfully constructed, which can be used as whole-process positive control for NiV nucleic acid extraction and detection.